Biotin Rat Anti-Mouse CD25
Clone 7D4 (RUO)
- Brand BD Pharmingen™
- Alternative Name IL-2 Receptor α chain, p55
- Concentration 125 µg/ml
- Isotype Rat LEW, also known as Lewis IgM, κ
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
Immunohistochemistry-frozen (Tested During Development)
Immunohistochemistry-paraffin (Not Recommended)
- Immunogen IL-2-dependent BALB/c mouse helper T-cell clone HT-2
- Storage Buffer Aqueous buffered solution containing BSA, goat serum, and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The 7D4 monoclonal antibody specifically binds to CD25, the low affinity IL-2 Receptor (IL-2R α , p55) expressed on activated T and B lymphocytes from all mouse strains tested. IL-2R α by itself is not a signaling receptor. However, it can combine with IL-2 Receptor β (CD122) and γ (CD132) chains to form high-affinity, signaling receptor complexes for IL-2. Resting T and B lymphocytes and resting and activated NK cells do not express IL-2R α . CD25 is transiently expressed at a low level during normal B-cell development in the bone marrow on the CD45R/B220low TdT- sIg- Pre-B/Pre-B-II and CD45R/B220low TdT- sIgM+ sIgD- immature B stages, but not on the CD45R/B220low TdT+ sIg- Pro-B/Pre-B-I stage nor on CD45R/B220high TdTsIgM+ sIgD+ mature B cells. It is expressed at a higher level during a very early stage of T-cell development in fetal and adult thymus. Peripheral CD25+ CD4+ T lymphocytes called regulatory T (Treg) cells are involved in the maintenance of self-tolerance. It has also been reported that dendritic cells express CD25, recognized by mAb 7D4. The 3C7 antibody recognizes an epitope of CD25 which is distinct from those recognized by mAbs 7D4 and PC61, and it blocks binding of IL-2 to CD25.
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Preparation and Storage
Store undiluted at 4°C.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Use of these products to measure activation antigens expressed on mononuclear cell subsets for the purpose of monitoring immunoregulatory status can fall under one or more claims of the following patents: US Patent Nos. 5,445,939, 5,656,446, 5,843,689; European Patent No. 319,543; Canadian Patent No. 1,296,622; Australian Patent No. 615,880; and Japanese Patent No. 2,769,156.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- This antibody has been developed for the immunohistochemistry application. However, a routine immunohistochemistry test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
Immunohistochemistry: The 7D4 antibody specific for mouse CD25 is recommended to test for immunohistochemical staining of acetone-fixed frozen sections. Tissues tested were mouse spleen and thymus. The antibody stains activated T and B lymphocytes and immature thymocytes. The isotype control recommended for use with this antibody is biotin-conjugated rat IgM (Cat. No. 550343). For optimal indirect immunohistochemical staining, the biotin format of the 7D4 antibody should be titrated (1:10 to 1:50 dilution) and visualized by Streptavidin-HRP (Cat. No. 550946) together with the DAB detection system (Cat. No. 550880). A detailed protocol of the immunohistochemical procedure can be found on our website at www.bdbiosciences.com/support/resources . The clone 7D4 is not recommended for formalin-fixed paraffin embedded sections.